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OBJECTIVE Alzheimer's disease(AD)is the most common neurodegenerative disease worldwide.Neuroinflammation is a potential target for the patients with AD.It is attributed to activated microglia and the release of various inflammatory mediators from infec-tion,ischemia and toxin accumulation.Accumulating evi-dence has indicated that the cGAS-STING pathway driven neuroinflammation in neurological disease.TSG is a main natural active ingredient that derived from polyg-onum multiflorum.Previous research from our group found that TSG has beneficial effects of anti-aging,anti-inflammatory action and improving memory function in APP/PS1 transgenic AD mice.Here,we investigated the effects of TSG on cognitive impairment and neuroinflam-mation in APP/PS1-AD mice and explore the underly-ing mechanism by which TSG ameliorates memory func-tion in the cGAS-STING-mediated inflammatory response.METHODS The Morris water mace test and the novel object recognition test were performed to test the effects of TSG on spatial learning and cognitive and memory abil-ity in APP/PS1 double transgenic AD mice model.In addi-tion,real-time quantitative PCR,Western blotting,ELISA analysis,and flow cytometry to examine gene and pro-tein expression of cGAS-STING related pro-inflammatory cytokines and chemokines.Statistical analyses were ana-lyzed using the SPSS 25.0 package by analysis of vari-ance(ANOVA).Neuman-Keuls or Tukey's multiple-com-parisons test were conducted as ANOVA justified post hoc comparisons between group means.RESULTS We demonstrated that AD transgenic mice exhibited cognitive deficits accompanied by the elevated serum and brain inflammation.The expressions of serum inflammatory cytokines and the activation of microglia in cerebral cor-tex and hippocampus were suppressed after TSG treat-ment,which was probably attributable to the decrease of cyclic GMP-AMP synthase(cGAS)and stimulator of interferon genes(STING)triggered immune response.Additionally,the data showed that TSG treatment reduced the expression level of inflammatory cytokines(IL-1β,TNF-α,IFN-β,IFN-α)in microglial cells BV2 primed with LPS and IFN-γ.CONCLUSION TSG implicated the health benefits in preventing cognitive disorders by inhib-iting neuroinflammation via cGAS-STING signalling path-way in AD.
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OBJECTIVE：To study the regulatory effects of stilbene glucosid e（TSG）on c-Jun N-terminal kinase （JNK）and protein phosphortase 2B（PP2B）in APP/PS1/Tau transgenic dementia （3×Tg-AD）mice，and to explore its potential mechanism of anti-Alzheimer’s disease （AD）. METHODS ：Totally 45 male 3×Tg-AD mice were randomly divided into model group ，positive control group （huperzine A ，0.15 mg/kg），TSG low-dose ，medium-dose and high-dose groups （0.033，0.1，0.3 g/kg），with 9 mice in each group. Another 9 normal male C 57BL/6J mice were included into normal control group. Administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 60 d. Normal control group and model group were given constant volume of normal saline intragastrically. After medication ，Morris water maze experiment was used to test the spatial learning and memory ability of mice in each group ；Nissl staining was used to observe the changes of Nissl bodies in cerebral cortex and hippocampus ；mRNA and protein expressions of JNK and PP 2B were detected by qRT-PCR and Western blotting assay. RESULTS：Compared with normal control group ，the escape latency was significantly prolonged （P＜0.01），the retention time of the original platform quadrant was significantly shortened （P＜ and the times of crossing the platform was significantly reduced in model group （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was significantly 729011126@qq.com reduced，the staining was slight ；the relative expressions of JNK mRNA and protein were significantly increased （P＜ 0.01），and the relative expressi ons of PP 2B mRNA and protein were significantly decreased （P＜0.01）. Compared with model group ，the escape latency was significantly shortened in positive control group and TSG groups （P＜0.01）；the retention time of the original platform quadrant was significantly prolonged （P＜0.01）；the times of crossing the platform was significantly increased （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was increased significantly ，the staining was heavy ；the relative expression of JNK protein was significantly decreased（P＜0.05 or P＜0.01），the relative expressions of PP 2B mRNA and protein were significantly increased （P＜0.01）， while the relative expression of JNK mRNA was significantly decreased in TSG high-dose group （P＜0.05）. CONCLUSIONS ：TSG can improve the learning and memory ability and neuronal damage of 3 × Tg-AD mice. The mechanism may be related to down-regulating the transcription and expression of protein kinase JNK ，up-regulating the transcription and expression of protein phosphatase PP 2B.
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OBJECTIVE：To study the effects of stilbene glucoside （TSG）on the proliferation and estrogen receptor （ER）of human breast cancer T- 47D cells ，and to explore its estrogen-like effect and potential mechanism. METHODS ：Taking ER positive human breast cancer T- 47D cells as subjects ，using β-estradiol（β-E2，1×10－8 mol/L）as positive control ，CCK-8 assay was used to detect the cell proliferation after treated with different concentrations of TSG （1×10－8，1×10－7，1×10－6，1×10－5，1×10－4 mol/L）for 24，48，72 h；the cell proliferation rate was calculated. Western blotting assay and RT-PCR methods were adopted to detect the protein and mRNA expression of ER-α and ER-β in cells after treated with low，medium and high concentrations of TSG （1×10－8， 1×10－6，1×10－4 mol/L）for 48 h. RESULTS ：After treated with different concentrations of TSG for 24，48，72 h，the cell proliferation rate of each administration group at each time point （except for β-E2 group at 48 h）increased significantly ，compared with blank group ；those of TSG groups （1×10－5，1×10－6，1×10－7 mol/L）were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. After treated with low ，medium and high concentrations of TSG for 48 h，protein and mRNA expression of ER-α and ER-β in cells were increased significantly，compared with blank group （P＜0.05 or P＜0.01）；protein expression of ER-β in TSG low concentration group ，mRNA expression of ER-α in TSG groups as well as mRNA expression of ER-β in TSG low and high concentration groups were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can induce the in vitro proliferation of T- 47D cells and exert estrogen-like effects by promoting protein and mRNA expression of ER-α and ER-β， which is stronger than that of β-E2 at a certain concentration.
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Objective: To investigate the neuroprotective effect and mechanism of tetrahydroxy stilbene glucoside (TSG) on β-amyloid protein 25-35 (Aβ25-35)-induced neuron synapses damage. Method: Primary neurons were isolated and purified from cerebral cortex of suckling mouse. Then neurons were divided into control group, model group (incubation with Aβ25-35) and TSG groups (after incubation with Aβ25-35, add 6.25, 12.5, 25, 50, 100 μmol·L-1 TSG). Cell counting kit-8 (CCK-8) and Lactate dehydrogenase (LDH) methods were used to observe the viability of neuron, immunocytochemical staining was performed to determine the expressions of synapsin-1 (SYN-1), and the concentration of postsynaptic density-95 (PSD-95) and synaptophysin (SYP) were detected by enzyme-linked immunosorbent assay (ELISA) method. The level of cyclic adenosine monophosphate response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) and the level of CREB, Phosphorylated CREB (p-CREB) and BDNF proteins were determined by immunocytochemical staining or Western blot (WB). Result: Compared with normal group, the cell survival rate of model group was significantly reduced, LDH release was significantly increased (PPPPPPP-1,25 μmol·L-1 TSG can significantly enhance the expression of SYN-1(PPPPConclusion: TSG possesses the neuroprotective effect on Aβ25-35-induced neuron synapses, the mechanism may be associated with the activation of CREB/BDNF signaling pathway.
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Objective:To optimize the high pressure steaming processing technology for Polygonum multiflorum Thunb.by Box-Behnken response surface methodology , and compare with the traditional processing .Methods:The effects of factors such as steaming temperature, steaming time and drying temperature on polysaccharide content , stilbene glucoside content and normalized value in Po-lygonum multiflorumThunb.were studied by Box-Behnken response surface methodology .The content differences of polysaccharides and stilbene glucoside between the high pressure steaming processed product and the traditional processed product were evaluated .Re-sul ts:The best high pressure steaming processing conditions for Poyl gonumm luitlfo urm Thunb .were as follows:the steaming tempera -ture was 1254.℃ , the steaming time was3.1 h, and the drying temperature was 52℃.The contents of polysaccharides and stilbene glycosides in Polygonum multiflorum Thunb.processed by the high pressure steaming method were 1.24-fold and 5.26-fold higher than those processed by the traditional method .Conclusion:Box-Behnken response surface method can be used to optimize the high pres-sure steaming processing for Polygonum multiflorum Thunb., and the method is simple and predictable .
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To explore the active substance of antiplatelet aggregation of Polygoni Multiflori Radix by using chemical fingerprints and antiplatelet aggregation bioactivity test for spectrum-effect correlation analysis. The Polygoni Multiflori Radix was tested by antiplatelet aggregation in vitro, and the results showed that 50% aqueous ethanol extract of Polygoni Multiflori Radix had more potent antiplatelet aggregation effect than 10% or 90% aqueous ethanol extract, and ultrasonic extraction was superior to refluxing extraction in the aspect of antiplatelet aggregation. The antiplatelet aggregation bioactivity of the different Polygoni Multiflori Radix extracts was evaluated and the results showed that the inhibition rate was 32.03%-74.56%. Spectrum-effect correlation analysis indicated that trans-stilbene glucoside, cis-stilbene glucoside and catechinic acid had higher correlation coefficient and they were 0.963 (P<0.01), 0.902 (P<0.01) and 0.656 (P<0.05) respectively; furthermore, all of the above three compounds demonstrated significant antiplatelet aggregation bioactivities. Considering their content difference in Polygoni Multiflori Radix, we calculated the relative active contributions, and the results suggested that trans-stilbene glucoside was the main active substance of Polygoni Multiflori Radix in the aspect of antiplatelet aggregation in vitro.
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AIM To establish an HPLC method for determining the contents of three constituents in Polygoni multiflori Radix Praeparata in plasma of atherosclerosis rats.METHODS After the rats were intragastrically administrated with Polygoni multiflori Radix Praeparata CMC-Na solution,the plasma was collected.The HPLC analysis was carried out on a 30 ℃ thermostatic Hypersil C1s column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.03％ phosphoric acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.DAS2.0 software was applied to drawing concentration-time curves and calculating pharmacokinetic parameters.RESULTS Stilbene glucoside,emodin and physcion showed good linear relationships within the ranges of 61.25-6 125 μg/L (r =0.999 8),12.6-3 150 μg/L (r =0.999 3) and 24.1-6 030 μg/L (r =0.999 5),respectively.The method recoveries were 99.5％-105.8％ with the RSDs of 1.3％-3.3％,while the extraction recoveries were 87.2％-96.3％ with the RSDs of 3.2％-5.9％.The pharmacokinetic behaviors of three constituents all accorded with two-compartment model,but their contents in plasma were much lower than those in medicinal material.CONCLUSION The bioavailabilities of stilbene glucoside,emodin and physcion are relatively low in plasma of atherosclerosis rats,which may be related to constituents' intestinal absorption after intragastric administration with Polygoni multiflori Radix Praeparata.
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Objective To analyze the main chemical constituents and their contents in aqueous extract of Polygoni Multiflori Radix (PMR, root of Polygonum multiflorum), and to elucidate the effects of aqueous extract of PMR and its main constituents on the expression of the mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells. Methods The main chemical constituents and their content in aqueous extract of PMR were determined by HPLC. The cytotoxicity of aqueous extract of PMR and its main constituents on L02 cells was determined by MTT assay. The mRNA expression of CYP1A2, CYP2C9, and CYP2E1 in L02 cells were detected by quantitative real-time PCR. Results There were four main well-separated chromatographic peaks standing for tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside, emodin and physcion in aqueous extract of PMR. The content of thesecomponents in aqueous extract of PMR was (1.14 ± 0.03)%, (0.106 9 ± 0.001 6)%, (0.010 8 ± 0.000 9)%, (0.003 55 ± 0.000 19)%, respectively. The cytotoxicity of aqueous extract of PMR and emodin on L02 cells at 24 h was dose-dependent, and the concentration of 50% inhibition was 7.290 mg/mL and 0.082 mmol/L respectively. Tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside and physcion did not show significant cytotoxicity on L02 cells in the experimental concentrations. Aqueous extract of PMR and emodin significantly inhibited the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells. Emodin-8-O-β-D-glucoside inhibited the expression of mRNA of CYP1A2 and CYP2C9. Tetrahydroxy stilbene glucoside inhibited the expression of mRNA of CYP1A2 but activated the expression of mRNA of CYP2C9. Physcion inhibited the expression of mRNA of CYP1A2 and CYP2C9 in a dose-dependent manner, but inhibited the expression of mRNA of CYP2E1 in low concentration and activitated the expression of mRNA of CYP2E1 in high concentration. Conclusion The inhibition of aqueous extract of PMR on the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells is the combined effect of all components in it. The main four components all inhibit the expression of mRNA of CYP1A2. The anthraquinone is the main component inhibiting the expression of mRNA of CYP2C9. The free anthraquinone is the main component inhibiting the expression of mRNA of CYP2E1.
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Stilbene glucoside is one of major active constitutions in Polygoni Multiflori Radix. Stilbene glucoside contributed markedly to resisting caducity, protecting nerve, reducing serum lipids, and protecting liver and antitumor activity. The extraction and purification technology is the key issue to obtain stilbene glucoside. Through research, extraction and purification technology has made a certain progress. The stilbene glycoside could be extracted by water boiling, diacolation, and backflow; then it could be purified by the method of chromatography, recrystallization, and extraction. In this article, the technologies in extracting and purifying stilbene glucoside were reviewed, and the prospect of stilbene glucoside preparation was discussed to lay the foundation for further research and application.
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Objective: To optimize a technology of integration on field processing and processing crude drugs of Polygoni Multiflori Radix (PMR) and to provide the scientific evidence for the integration of PMR. Methods: Orthogonal test was used to optimize the integration technology on primary processing and reprocessing with two major characteristic components (stilbene glucoside and combined anthraquinone). To compare the differences between the integration technology and traditional technology on pharmacological activity by models of constipation and swelling of ear in mice with Various index (small intestinal propulsion rate, first defecation time, fecal water content, swelling degree of ear, and inflammatory factors). Results: The results showed that the integration technology of 50℃, 16 h was better, and there was significant difference in chemical composition and Laxative effect between the integration technology and traditional technology. The anti-inflammatory effect of the integrated process was better than that of the traditional technology. Conclusion: The technology of integration of field processing and processing crude drugs is feasible and the operation is good.
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Objective: To study the effect of different processing methods and conditions for bioactive constituents in Polygoni Multiflori Radix (the root tubers of Polygonum multiflorum) and their change regularity, in order to optimize and establish appropriate drying methods and conditions. Methods: The stilbene glucoside, free-anthraxquinones, combined-anthraxquinones, and catechin from 12 kinds of Polygoni Multiflori Radix in different processing methods were determined by MEKC-DAD. The multiple functional substances of principal component analysis (PCA) was made to standardized treatment for the comprehensive evaluation of the 12 samples by different drying methods. Results: The contents of multiple functional substances in different processed products from Polygoni Multiflori Radix presented the certain regularity. The contents of stilbene glucoside and combined anthraquinone were the highest in Polygoni Multiflori Radix which was cut into thick pieces dried in the sun, the total free anthraquinones were the highest in Polygoni Multiflori Radix which was cut into thick pieces dried at 70℃, the content of catechin was the highest in Polygoni Multiflori Radix which was cut into thin pieces dried at 40℃. The comprehensive evaluation index obtained with PCA showed that the sample cut into thick pieces dried in the sun was significantly higher than those with other samples. Conclusion: The influence of processing methods by the determination of multiple functional substances in Polygoni Multiflori Radix has been investigated and a scientific basis for the suitable processing method of Polygoni Multiflori Radix provided.
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Objective: To determine the optimal preparation conditions for Huangmai granules. Methods: Using single factor screening method and orthogonal design method, the extraction process and granulation process were optimized. Results:The optimal preparation process was as follows:the medicinal materials in the prescription were extracted 3 times with 8-fold amount of water with 2h for each time. The water extract was combined followed by vacuum concentration, and then dried to obtain dry extract. The powder of dry extract was evenly mixed with dextrin at appropriate amount, and the wet granules were prepared with the 40% ethanol followed by drying and size stabilization to obtain the products. Conclusion:The selected preparation process is scientific, reasonable and sta-ble, and suitable for the industrial production.
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OBJECTIVE: To study the absorption and metabolism of active compounds of Polygoni Multiflori Radix in L-02 cells and elucidate their chemical structures by liquid chromatography tandem-mass spectrometry. METHODS: The active compounds in Polygoni Multiflori Radix, 2,3,5,4'-tetrahydroxystilbene-2-O-γ-5-glucoside (TSG) and emodin (EN), were cultured into L-02 cells. Cell culture media and cell lysis were collected after 0,2,4,6,8,12,24,36 and 48 h respectively. Reversed-phase high-performance liquid chromatography (HPLC) was used to explore the possible absorption and metabolism. Liquid chromatography-tandem-mass spectrometry (LC-MS/MS) was used to identify the chemical structures of the metabolic products. RESULTS: TSG showed no obvious accumulation in L-02 cells. One metabolic product of TSG was detected by HPLC, and its concentration increased with lime after 8 h. This metabolite was identified as the glucuronidation of TSG by LC-MS/MS. In contrast, EN was partially accumulated in L-02 cells. Three isomers of glucuronidations and one sulfation of EN were also identified by LC-MS/MS. CONCLUSION: EN is easier to be absorbed and accumulated in L-02 cells than TSG. Phase II metabolic reactions play an important role in the metabolism of TSG and EN in cells. However, the accurate binding sites need to be further confirmed.
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OBJECTIVE: To establish a micellar electrokinetic chromatography with diode array detection (MEKC-DAD) method for simultaneous determination of seven components including stilbene glucoside, emodin, aloe-emodin, rhein, chrysophanol, physcion, and catechin in Polygoni Multiflori Radix, of different origins and commercial herbs. METHODS: Based on the mode of MEKC, an uncoated fused silica capillary (75 μm × 64.5 cm, 56 cm of effective length) was adopted, and 25 mmol · L-1 borax-30 mmol · L-1 SDS-10% acetonitrile (pH 9.60) was selected for the running buffer. The detection wavelength was set at 254 nm. The separation voltage was 25 kV and the column temperature was maintained at 24℃. The sample was injected at 5 kPa for 5 s. RESULTS: The calibration curves of the seven components in Polygoni Multiflori Radix showed good linearity (r ≥ 0.9994). The average recoveries of the method were between 99.164%-101.504%. CONCLUSION: The established method is reliable and accurate, and can be used for the quality control of Polygoni Multiflori Radix.
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Objective: To optimize the percolation extraction technology for Qixue Shuangbu Tinctura by response surface methodology (RSM). Methods: The single factor experiment combined with Box-Behnken design was used to optimize the extracting technology, with six major characteristic components (ginsenoside Rb1, calycosin 7-O-β-D-glucopyranoside, ferulic acid, stilbene glucoside, paeoniflorin, hesperidin) and extract of Qixue Shuangbu Tinctura as indexes, in order to detect three factors, including the alcohol volume fraction, the dosage of ethanol, and the percolate speed, and optimize the extraction process of Qixue Shuangbu Tinctura. Results: Optimum percolation process was added 10 times of the amount of 48% ethanol with percolation speed of 1.4 mL/min. Conclusion: This optimized extraction technology of Qixue Shuangbu Tinctura is reasonable, stable, and feasible, and with high accuracy, which could be extended to the applications of large-scale production.
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Objective To screen and clone the genes related to stilbene glucosides biosynthesis in Polygonum multiflorum Thunb. Methods The differentially expressed genes in the root, stem and leaf of Polygonum multiflorum which have different contents of stilbene glucosides were screened by differential display reverse transcription polymerase chain reaction (DDRT-PCR). After pMD19-T carrier was inserted into the obtained differential genes for sequencing and comparison, the gene function was analyzed. Results Fifty-one differentially expressed cDNA fragments were found. Of them, 9 were used for the identification by semi-quantitative PCR. The identification results presented 3 positive fragments, one fragment was specifically expressed in the stem and leaf of Polygon-um multiflorum Thunb., sharing high homology with glycine dehydrogenase, and 2 were specifically expressed in the root of Polygonum multiflorum Thunb., having high homology with enoyl-CoA hydratase and aminopeptidase N, respectively. Conclusion Three homologous gene sequences obtained through DDRT-PCR provide a basis for the further study of biosynthetic pathway of stilbene glucoside from Polygonum multiflorum Thunb..
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@#Objective To investigate the deficit of extracellular-signal regulated kinase (ERK)1/2 activation in the different age of Alzheimer's disease (AD)-like animal model and the protective effect of 2,3,5,4′-tetrahydroxy stilbene-2-O-β-D-glucoside(TSG), which is the main component of Polygonum multiflorum, on ERK activation. Methods A generally accepted animal model of AD - PDAPPV717I transgenic (Tg) mouse was observed from 4 to 16 months old. Tg mice were randomly divided into 3 model groups(4, 10 and 16 months old mice)and TSG treated (at doses 120 and 240 μmol/kg/d) groups. TSG was administered to some Tg mice with an age range 4-10 months. In untreated 10 months old Tg mice, the TSG was administrated to those falling in the age range 10-16 months. For the control group we adopted the same age and background C57BL/6J mice. The ERK1/2 expression and phosphorylation were detected by Western blotting.Results In the 4-month-old PDAPPV717I Tg mice, phosphorylation of ERK1/2 decreased significantly in hippocampus and cortex compared with age matched control. In the 10-month-old Tg mice, decrease of ERK1/2 activation was aggravated in cortex but was less in hippocampus. The treatment of TSG at the doses of 120 and 240 μmol/kg for 6 months (from the age of 4 to 10 months) significantly up-regulated ERK1/2 activation in Tg mice. In the 16-month-old Tg mice, over-activation of ERK1/2 occurred in both hippocampus and cortex. The transgenic mice treated by TSG for 6 months (from the age of 10 months to 16 months) showed significant inhibition of over-activation of ERK1/2. Expression of total ERK1/2 showed no difference among control, Tg model and TSG treated groups.Conclusion PDAPPV717I transgenic mice with an age range from 4 to 16 months revealed the time-dependent deficit of ERK1/2 activation. TSG can bring the down or over activation of ERK1/2 into normal. Because ERK1/2 activation plays the crucial role in cellular signal transduction and learning-memory ability, TSG may have beneficial potential to the prevention and treatment of neurodegenerative diseases like AD.
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OBJECTIVE: To establish the quality standard of Yishen granules.METHODS:Polygonum multiflorum,Ganoderma,Radix Astragali and Angelica sinensis in the formulation were identified by TLC.The content of stilbene glucoside in polygonum multiflorum was determined by HPLC.RESULTS: The TLC identification method exhibited good specificities.The samples were similar to the references /control in that their TLC spots assumed identical color at the corresponding places,yet without the interference form the negative samples.The linear range of stilbene glucoside was 0.037 75～0.755 ?g(r=0.999 5) and the average recovery was 99.44%(RSD=1.79%,n=6).CONCLUSION: The established standard is accurate,reliable and applicable for the quality control of Yishen granules.
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Objective To determinate the content of 2, 3, 5, 4′-stibene glucoside in Xuezhiping Capsules. Method HPLC was performed with Diamosil~(TM)C_(18) column, a mobile phase of acetonitrile-water (20: 80), the detection wavelength at 320nm and the flow rate being 1.0 mL/mL. Results 2, 3, 5, 4′-stilbene glucoside has a good linear within 0.0267～2.6640 ?g and the average recovery was 99.62% with RSD 1.32%. Conclusion This method is convenient and efficient and can be used for the quality control of Xuezhiping Capsules.
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Objective To provide basis for choosing a proper processed method of Polygonum multiflorum.Methods To compare and analyze main active component changes in the processed products of P.multiflorum by being steamed without any adjuvants and with soya-bean milk of different concentrations.Results There was no marked difference for variously processed products in the main active components,anthraquinone and stilbene glucoside in P.multiflorum.Conclusion It is feasible to replace the steamed with soya-bean milk by the steamed without any adjuvants.